Servicio de Información Comunitario sobre Investigación y Desarrollo - CORDIS

Primers and protocols for PCR amplification of selected genes whose expression is indicative for dormancy or hardiness in beech

We selected 4 beech genes that are likely to be involved in beech cold hardiness development and developed RT-PCR primers to allow rapid and robust analysis of the quantitative expression levels of these genes. It concerns
- ABA inducible protein
- alpha tubulin
- a gene with unknown function
- a protein kinase

These primers can be used for quantitative analysis of stress related gene expression.
The can be used in scientific research.
In addition, they may serve as content for a molecular diagnostic assay for hardiness of beech seedlings
We developed a reliable protocol that takes into account the difficulties of the tissue type (dormant buds). The protocol starts with a CTAB RNA isolation. Preparations were DNAseI and used for cDNA synthesis. Dilutions of this cDNA were used for Realtime PCR. Product formation was measured using the iCycler system (BIORAD Laboratories, The Netherlands). Relative changes in expression were calculated using the Gene Expression Macro (Version 1.1) supplied by BIORAD.

We will use these primers in follow up research projects and for the development of diagnostics tests

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Agrotechnology & Food Innovations BV
P.O. Box 17
6700 AA Wageningen
Netherlands
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